• First do pixel clustering, then do cell clustering using frequency of pixel clusters
  • Gaussian blur sigma=2, k=20 (pixel metaclusters)
  • For each pixel, divide each marker by total (each pixel sums to 1)
  • z-score cap=3 for hierarchical clustering
  • k=20 (cell metaclusters)

Rep 1

FlowSOM

Heatmap of pixel metacluster x cell cluster

Heatmap of marker x cell cluster (integrated)

Heatmap of marker x cell cluster (weighted)

Rep 2

FlowSOM

Heatmap of pixel metacluster x cell cluster

Heatmap of marker x cell cluster (integrated)

Heatmap of marker x cell cluster (weighted)

Rep 3

FlowSOM

Heatmap of pixel metacluster x cell cluster

Heatmap of marker x cell cluster (integrated)

Heatmap of marker x cell cluster (weighted)

Rep 4

FlowSOM

Heatmap of pixel metacluster x cell cluster

Heatmap of marker x cell cluster (integrated)

Heatmap of marker x cell cluster (weighted)

Rep 5

FlowSOM

Heatmap of pixel metacluster x cell cluster

Heatmap of marker x cell cluster (integrated)

Heatmap of marker x cell cluster (weighted)